Chen Andrew L, Fang Carrie, Liu Chuanju, Leslie Michael P, Chang Eric, Di Cesare Paul E
Department of Orthopaedic Surgery, Musculoskeletal Research Center, NYU-Hospital for Joint Diseases Orthopaedic Institute, 301 East 17th Street, New York, NY 1003, USA.
J Orthop Res. 2004 Nov;22(6):1188-92. doi: 10.1016/j.orthres.2004.02.013.
Coordinate expression of BMPs and their receptors and inhibitors is likely necessary for physiologic BMP regulation and activity. To characterize the expression of such factors in fetal, normal adult, and end-stage osteoarthritic articular cartilage, samples from these sources were analyzed. PCR-amplified sequences (BMPs 1-11), receptors (IA, IB, II), TGF-beta1, TGF-beta2, inhibitors noggin and follistatin, CDMP-1, COMP, and GAPDH from cDNAs generated from extracted total RNA were resolved by gel electrophoresis. Protein levels of BMPs 3, 7, and 8 were also analyzed by SDS-PAGE and Western blotting. RT-PCR revealed that BMPs 1, 2, 4-6, and 11, BMPR-IA and II, noggin, follistatin, CDMP-1, COMP, and GAPDH mRNAs were expressed in similar fashion in both fetal and adult (normal or osteoarthritic) cartilage. BMPs 9 and 10 mRNAs were not expressed in either group. BMPs 7, 8, and BMPR-IB mRNAs were consistently expressed in fetal but not in adult cartilage. BMP-3 mRNA was expressed in fetal and normal adult, but not in osteoarthritic samples. TGF-beta1 was expressed in both adult normal and osteoarthritic, but not fetal, samples. Similarly, Western blotting demonstrated BMPs 7 and 8 to be present in fetal but not in adult samples. BMP-3 protein was present in fetal and adult normal samples, to a lesser extent, but absent in osteoarthritic cartilage.
骨形态发生蛋白(BMPs)及其受体和抑制剂的协同表达可能是生理状态下BMP调节和活性所必需的。为了表征这些因子在胎儿、正常成人及终末期骨关节炎关节软骨中的表达情况,对来自这些来源的样本进行了分析。通过凝胶电泳解析了从提取的总RNA生成的cDNA中的PCR扩增序列(BMPs 1 - 11)、受体(IA、IB、II)、转化生长因子-β1(TGF-β1)、转化生长因子-β2(TGF-β2)、抑制剂头蛋白(noggin)和卵泡抑素(follistatin)、软骨分化相关蛋白-1(CDMP-1)、软骨寡聚基质蛋白(COMP)以及甘油醛-3-磷酸脱氢酶(GAPDH)。还通过SDS-PAGE和蛋白质印迹法分析了BMPs 3、7和8的蛋白质水平。逆转录-聚合酶链反应(RT-PCR)显示,BMPs 1、2、4 - 6和11、骨形态发生蛋白受体-IA(BMPR-IA)和II、头蛋白、卵泡抑素、CDMP-1、COMP以及GAPDH的信使核糖核酸(mRNAs)在胎儿和成人(正常或骨关节炎)软骨中的表达方式相似。BMPs 9和10的mRNAs在两组中均未表达。BMPs 7、8和骨形态发生蛋白受体-IB(BMPR-IB)的mRNAs在胎儿软骨中持续表达,但在成人软骨中不表达。BMP-3的mRNAs在胎儿和正常成人软骨中表达,但在骨关节炎样本中不表达。TGF-β1在成人正常和骨关节炎样本中表达,但在胎儿样本中不表达。同样,蛋白质印迹法表明BMPs 7和8存在于胎儿样本中,但不存在于成人样本中。BMP-3蛋白存在于胎儿和成人正常样本中,但含量较少,在骨关节炎软骨中不存在。